Abstract
In this protocol, we provide a detailed step-by-step bacterial surface imaging and molecular analysis procedure. With SPM (scanning probe microscopy)-based dynamic force microscopy (DFM) imaging, we achieved a so far unprecedented resolution of ~1 nm on the outer surface layer of Tannerella forsythia and monitored the production of curli fibers on Escherichia coli in physiological conditions. Moreover, using these immobilization methods, single-molecule force spectroscopy experiments were conducted on living bacterial cells.
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References
Engel A, Muller DJ (2000) Observing single biomolecules at work with the atomic force microscope. Nat Struct Biol 7(9):715–718. https://doi.org/10.1038/78929
Martinez-Martin D, Herruzo ET, Dietz C, Gomez-Herrero J, Garcia R (2011) Noninvasive protein structural flexibility mapping by bimodal dynamic force microscopy. Phys Rev Lett 106(19):198101
Rico F, Su C, Scheuring S (2011) Mechanical mapping of single membrane proteins at submolecular resolution. Nano Lett 11(9):3983–3986. https://doi.org/10.1021/nl202351t
Arnoldi M, Fritz M, Bauerlein E, Radmacher M, Sackmann E, Boulbitch A (2000) Bacterial turgor pressure can be measured by atomic force microscopy. Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics 62(1 Pt B):1034–1044
Kuznetsova TG, Starodubtseva MN, Yegorenkov NI, Chizhik SA, Zhdanov RI (2007) Atomic force microscopy probing of cell elasticity. Micron 38(8):824–833. https://doi.org/10.1016/j.micron.2007.06.011
Mortensen NP, Fowlkes JD, Sullivan CJ, Allison DP, Larsen NB, Molin S, Doktycz MJ (2009) Effects of colistin on surface ultrastructure and nanomechanics of Pseudomonas aeruginosa cells. Langmuir 25(6):3728–3733. https://doi.org/10.1021/la803898g
Touhami A, Nysten B, Dufrêne YF (2003) Nanoscale mapping of the elasticity of microbial cells by atomic force microscopy. Langmuir 19(11):4539–4543. https://doi.org/10.1021/la034136x
Chtcheglova LA, Waschke J, Wildling L, Drenckhahn D, Hinterdorfer P (2007) Nano-scale dynamic recognition imaging on vascular endothelial cells. Biophys J 93(2):L11–L13. https://doi.org/10.1529/biophysj.107.109751
Florin EL, Moy VT, Gaub HE (1994) Adhesion forces between individual ligand-receptor pairs. Science (New York, NY) 264(5157):415–417
Lee GU, Chrisey LA, Colton RJ (1994) Direct measurement of the forces between complementary strands of DNA. Science (New York, NY) 266(5186):771–773
Hinterdorfer P, Baumgartner W, Gruber HJ, Schilcher K, Schindler H (1996) Detection and localization of individual antibody-antigen recognition events by atomic force microscopy. Proc Natl Acad Sci 93(8):3477–3481
Benoit M, Gabriel D, Gerisch G, Gaub HE (2000) Discrete interactions in cell adhesion measured by single-molecule force spectroscopy. Nat Cell Biol 2(6):313–317. https://doi.org/10.1038/35014000
Oh Yoo J, Cui Y, Kim H, Li Y, Hinterdorfer P, Park S (2012) Characterization of Curli a production on living bacterial surfaces by scanning probe microscopy. Biophys J 103(8):1666–1671. https://doi.org/10.1016/j.bpj.2012.09.004
Oh YJ, Hubauer-Brenner M, Gruber HJ, Cui Y, Traxler L, Siligan C, Park S, Hinterdorfer P (2016) Curli mediate bacterial adhesion to fibronectin via tensile multiple bonds. Sci Rep 6:33909. https://doi.org/10.1038/srep33909
Oh YJ, Sekot G, Duman M, Chtcheglova L, Messner P, Peterlik H, Schäffer C, Hinterdorfer P (2013) Characterizing the S-layer structure and anti-S-layer antibody recognition on intact Tannerella forsythia cells by scanning probe microscopy and small angle X-ray scattering. J Mol Recognit 26(11):542–549. https://doi.org/10.1002/jmr.2298
Allison DP, Mortensen NP, Sullivan CJ, Doktycz MJ (2010) Atomic force microscopy of biological samples. Wiley Interdiscip Rev Nanomed Nanobiotechnol 2(6):618–634. https://doi.org/10.1002/wnan.104
Doktycz MJ, Sullivan CJ, Hoyt PR, Pelletier DA, Wu S, Allison DP (2003) AFM imaging of bacteria in liquid media immobilized on gelatin coated mica surfaces. Ultramicroscopy 97(1–4):209–216
Dufrêne YF (2002) Atomic force microscopy, a powerful tool in microbiology. J Bacteriol 184(19):5205–5213. https://doi.org/10.1128/jb.184.19.5205-5213.2002
Baumgartner W, Hinterdorfer P, Schindler H (2000) Data analysis of interaction forces measured with the atomic force microscope. Ultramicroscopy 82(1):85–95
Zhu R, Gruber HJ, Hinterdorfer P (2018) Two ligand binding sites in Serotonin transporter revealed by nanopharmacological force sensing. Nanoscale imaging: Methods Mol Biol Vol.1814
Puntheeranurak T, Neundlinger I, Kinne RKH, Hinterdorfer P (2011) Single-molecule recognition force spectroscopy of transmembrane transporters on living cells. Nat Protocols 6(9):1443–1452
Acknowledgments
This work was supported by an APART (Austrian Programme for Advanced Research and Technology) fellowship of the Austrian Academy of Science.
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Oh, Y.J., Hinterdorfer, P. (2018). Sensing the Ultrastructure of Bacterial Surfaces and Their Molecular Binding Forces Using AFM. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_21
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DOI: https://doi.org/10.1007/978-1-4939-8591-3_21
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